Figure 1: Influence of the 3’RR deletion on IgG1 CSR and secretion.
From: Elucidation of IgH 3′ region regulatory role during class switch recombination via germline deletion
(a) ELISA analysis of IgG1 in supernatants of 3 days LPS+IL4 cultured B-cells. Data are the mean±s.e.m. of 8 experiments with 1 mouse (Mann–Whitney U-test for significance). (b) B splenocytes of wt and 3′RR-deficient mice were isolated (upper panel) and stimulated with LPS+IL-4 for 3 days (lower panel). Cells were labelled with anti-B220-APC antibodies and anti-IgG1-FITC antibodies. Percentage of B220+IgG1+ cells is reported in the gate. One representative experiment out of four (one mouse per experiment) is shown. (c) Southern blot analysis of Sμ-Cγ1 junctions amplified by PCR and hybridized with a 5′Cγ1 probe from 3 days LPS+IL4 stimulated splenocytes from wt, 3’RR-deficient mice and AID-deficient mice. DNA from AID-deficient B-cells (devoid of any CSR) was used as germline control DNA. Fifty nanograms and 20 ng of DNA were used for PCR experiments, respectively. One representative experiment out of four (one mouse per experiment) is shown. (d) Real-time PCR analysis of post switched IgG1 transcripts in 3 days LPS+IL4 cultured B cells. Values were normalized to Gapdh transcripts. Data are the mean±s.e.m. of 8 to 11 independent experiments with one mouse (Mann–Whitney U-test for significance). (e) ELISA analysis of IgG1 in 12-week-old mice sera. Data are the mean±s.e.m. of 8 mice for each genotype (Mann–Whitney U-test for significance).
